Non-enzymatic Mechanisms Research Articles

ATR-FTIR characterisation of daily-use plastics mycodegradation

Synthetic polymers, such as plastics, have permeated all aspects of modern life, and nowadays plastic pollution is a major environmental problem. Mycodegradation of these polymers could represent part of the solution to this problem since it calls on a broad toolbox of enzymes and applies non-enzymatic mechanisms to degrade and deteriorate recalcitrant materials. However, not enough is known about this ability for most of the representatives of the fungal kingdom. Another bottleneck is the harmonisation of technologies to analyse plastic degradation. This work involved the design of a biodegradation experiment, where the potential of four fungi representative of Dikarya and Penicillia (Funalia floccosa, Trametes versicolor, Pycnoporus cinnabarinus and Penicillium oxalicum) were tested on their ability to deteriorate the six most used plastics based on gravimetry and attenuated total reflectance-Fourier transform infrared spectroscopy (ATR-FTIR). The following correlation between changes in the band signals and the loss of mass after treatment was determined using polyethylene terephthalate, polypropylene, polyethylene, poly vinyl chloride, high density polyethylene, low density polyethylene and nylon. After treatment, the decrease in absorbance of the characteristic bands of the plastics was taken as an indication of the degradation of the corresponding bonds/functionalities. The four fungi used could transform CH, CH2, CH3, CO, CO, CN, NH and CCl bonds. The best result was obtained using the fungus F. floccosa with 90-day treatments for high density polyethylene (∼ 62.0 %), low density polyethylene (∼ 23.6 %) and nylon (∼ 35.6 %). Therefore, mycodegradation could open up new doors in the fight against plastic pollution.

Antiviral role of cholesterol 25-hydroxylase in inhibiting porcine circovirus 3 replication

Cholesterol 25-hydroxylase (CH25H) has significant antiviral effects through the production of 25-hydroxycholesterol (25HC). In this study, we investigated the effects of CH25H, its catalytic product 25HC, and its catalytic mutant lacking hydroxylase activity (CH25H-M) on porcine circovirus 3 (PCV3) replication. By transfecting PCV3 persistently infected PK-15 cells with the pCAGGS-CH25H-Flag plasmid, the results demonstrated that overexpression of CH25H significantly inhibited PCV3 Cap protein expression, Cap mRNA levels, and viral titers in a dose-dependent manner. Moreover, its catalytic product 25HC inhibited PCV3 replication in PK-15 cells at concentrations below 10 µM without affecting cell viability. In contrast, knockdown of endogenous CH25H using small interfering RNA (siRNA) enhanced PCV3 replication, further confirming its antiviral role. Interestingly, the CH25H-M mutant also exhibited inhibitory effects on PCV3 replication, although the inhibition was much less effective compared with CH25H. In conclusion, CH25H plays a critical role in regulating PCV3 replication, and its antiviral effect is not entirely dependent on its enzymatic activity. These findings provide new insights into both the enzymatic and non-enzymatic antiviral mechanisms of CH25H and revealed some mechanistic immune evasion for PCV3.

Metabolomics analysis reveals the non-enzymatic browning mechanism of green peppers (Piper nigrum L.) during the hot-air drying process

Non-enzymatic browning (NEB) reduced the colour quality of hot-air dried peppers, but the specific mechanism remains unclear. This may be related to the degradation of chlorophyll, ascorbic acid, and polyphenols. The findings revealed that the surface of pepper gradually browned during hot-air drying, with the ΔE⁎ value and browning degree (BD) significantly increasing by 119.4 % and 62.9 %, respectively. However, the total phenol content (TPC) and ascorbic acid content decreased by 74.2 % and 84.3 %, respectively. TPC was negatively correlated with BD (R = −0.86), and its value (31.7 %) was 300 times higher than that of other colour-related components (ascorbic acid, chlorophyll). UPLC-MS/MS analysis further identified 345 polyphenols, among which 1, 3-dicaffeoylquinic acid and 5,7-dihydroxy-3′, 4′, 5′-trimethoxyflavone were the two key monophenols influencing NEB (R = −0.87). The results suggested that NEB pathways involve chlorophyll and ascorbic acid degradation, the Maillard reaction, and polyphenol degradation, with the latter playing a major role.

Total Synthesis and Anticancer Evaluation of BZR-cotoxin IV

Anticancer peptides (ACPs) is a potential alternative for future cancer therapy. ACPs specifically inhibit cancer cells through a non-enzymatic membranolysis mechanism. One potential anticancer peptide worthy of investigation is BZR-cotoxin IV. Currently, research on BZR-cotoxin IV is limited to isolation, making it an interesting area for further development. One approach to developing BZR-cotoxin IV is through chemical synthesis techniques. This compound belongs to the group of cyclodepsipeptides that are naturally produced by the fungus Bipolaris zeicola Race 3. The synthesis of BZR cotoxin IV involves 3 steps: (1) Synthesis of the hydroxy acid precursor by conversion of L-valine amino acid to (2R)-hydroxyisovaleric acid, (2) synthesis of the linear depsipeptide using solid-phase peptide synthesis on 2-chlorotrityl chloride resin and (3) cyclization of the linear depsipeptide using solution phase synthesis to produce the BZR cotoxin IV compound with a purity of 11.7 %. BZR-cotoxin IV was characterized using HR-TOF-MS and 1H and 13C-NMR to validate the desired product. Anticancer activity testing was performed using the Resazurin assay on the HeLa cancer cell line, resulting in an IC50 of 187.50 μg/mL (214.014 μM) categorized as moderate. HIGHLIGHTS The chemical synthesis of BZR-cotoxin IV compounds has not been firstly reported. The synthesis of BZR-cotoxin IV was accomplished through a combination of solid-phase and solution-phase peptide synthesis. The objective was to synthesize and evaluate BZR-cotoxin IV for its cancer properties. GRAPHICAL ABSTRACT

Rapid Analysis for α-Tocopherol and Its Oxidative Products in the Pisum sativum L. Leaf Using Supercritical Fluid Chromatography-Medium Vacuum Chemical Ionization Tandem Mass Spectrometry.

A method for the rapid determination of α-tocopherol (α-T) and its oxidative products in plant tissue has been developed using supercritical fluid extraction (SFE) coupled with supercritical fluid chromatography (SFC) and medium vacuum chemical ionization (MVCI) with tandem mass spectrometry. The method is designed to study changes in levels for α-T and its oxidative products in plant cells during photosynthesis, aiming to observe the light response curves. α-T oxidation is a non-enzymatic self-defense mechanism in plant cells. Unlike enzyme-involved reactions, it cannot be stopped, so the oxidation continues in crude extracts even after extraction. Therefore, a real-time in-situ method is essential for tracking the light response curves. To optimize the selective reaction monitoring method, the reaction mixture of α-T and singlet oxygen (1O2), generated by rose Bengal under light illumination, was used as the source of oxidative products. The relative abundance changes in α-tocopherylquinone and 8a-hydroperoxy tocopherone in Pisum sativum L. (Pea) leaves under excessive light illumination have been preliminarily analyzed as part of the light response curve study. The method archives a throughput of 10-15 minutes for analyzing duplicate leaf samples. This process includes cutting off the leaf, sectioning it, placing the sample in a frozen SFE vessel, and conducting SFE/SFC analysis. Consequently, the average throughput is approximately 5-7 minutes per sample.

New Trichoderma Strains Suppress Blue Mold in Oranges by Damaging the Cell Membrane of Penicillium italicum and Enhancing Both Enzymatic and Non-Enzymatic Defense Mechanisms in Orange Fruits

Blue mold disease, caused by Penicillium italicum (P. italicum), presents a significant challenge to orange fruits (Citrus sinensis L.) and other citrus crops globally. Biological control, particularly Trichoderma species, offers a promising alternative to synthetic fungicides. Therefore, this study aimed to isolate, identify, and evaluate the antagonistic activities of two Trichoderma isolates against P. italicum. These isolates were molecularly identified and assigned accession numbers PP002254 and PP002272, respectively. Both isolates demonstrated significant antifungal activity in dual culture assays. Moreover, the culture filtrates (CFs) of Trichoderma longibrachiatum PP002254 and Trichoderma harzianum PP002272 suppressed the mycelial growth of P. italicum by 77.22% and 71.66%, respectively. Additionally, CFs reduced the severity of blue mold on orange fruits by 26.85% and 53.81%, compared to 100% in the control group. Scanning electron microscopy revealed that treated P. italicum hyphae were shrunken and disfigured. Enzyme activities (catalase, peroxidase, polyphenol oxidase, and phenylalanine ammonia-lyase) in treated oranges increased, along with total soluble phenolics and flavonoids. Conversely, malondialdehyde (MDA) levels decreased in treated fruits. These findings suggest that T. longibrachiatum PP002254 and T. harzianum PP002272 could be effective biocontrol agents for managing blue mold and other citrus postharvest diseases.

Ubiquitin ligase signalling networks shape presynaptic development, function and disease.

Ubiquitin ligases are important regulators of nervous system development, function and disease. To date, numerous ubiquitin ligases have been discovered that regulate presynaptic biology. Here, we discuss recent findings on presynaptic ubiquitin ligases that include members from the three major ubiquitin ligase classes: RING, RBR and HECT. Several themes emerge based on findings across a range of model systems. A cadre of ubiquitin ligases is required presynaptically to orchestrate development and transmission at synapses. Multiple ubiquitin ligases deploy both enzymatic and non-enzymatic mechanisms, and act as hubs for signalling networks at the synapse. Both excitatory and inhibitory presynaptic terminals are influenced by ligase activity. Finally, there are several neurodevelopmental disorders and neurodegenerative diseases associated with presynaptic ubiquitin ligases. These findings highlight the growing prominence and biomedical relevance of the presynaptic ubiquitin ligase network.

Improving grapevine (Vitis vinifera L., cv. Superior Seedless) drought tolerance with cerium oxide nanoparticles: Agronomic and molecular insights

Drought stress poses a significant threat to global food security, impacting grapevine growth and development through physiological, biochemical, and molecular alterations. Cerium oxide nanoparticles (CeO2 NPs) have recently gained attention as a potential solution to environmental challenges, yet their application in grapevines remains understudied. This study examined the interaction between foliar-applied CeO2 NPs (at concentrations of 0, 5, 50, and 500 mg L−1) and drought stress (at 30% and 70% of field capacity humidity) in grapevine saplings. Among the different concentrations tested, 50 mg L−1 CeO2 NPs significantly improved the agronomic traits (e.g., shoot length, leaf area, shoot and root dry weights), photosynthetic parameters (stomatal conductance, chlorophyll a and chlorophyll b) and RWC of the grapevine saplings under drought stress. In parallel, CeO2 NPs significantly induced the activity of compatible solutes and SOD, CAT and APX under drought stress conditions. Moreover, the availability of CeO2 NPs alleviated drought stress-induced damage in grapevine saplings, as evidenced by decreased H2O2 (32.63%), EL (40.35%) and MDA (50.63%) levels. The molecular results revealed that CeO2 NPs caused significant changes in gene expression under drought stress by reducing the expression of the VvCLH1, VvCu/ZnSOD, VvRD29A and VvRBOHA genes by 4.54-fold, 27.73-fold, 14.6-fold and 46.12-fold, respectively. These findings suggest that CeO2 NPs applied via leaves enhance grapevine sapling resistance to drought-induced oxidative stress through cooperative enzymatic and nonenzymatic antioxidant mechanisms, influencing gene regulation. As a result, these findings revealed that CeO2 NPs could be promising elicitor candidates for alleviating drought stress in grapevines.

Microbial interactions modify saffron traits selectively and modulate immunity through adaptive antioxidative strategy: Organic cultivation modules should be trait and crop-specific

Saffron (Crocus sativus L.) is a high-value, low-volume niche cash crop that has the unique ability to synthesize apocarotenoids. Since different biofertilizers are advocated for promoting plant health, we aimed to evaluate the interactions of commercially available microbial inoculants with this unique hysteranthous plant species. We studied the interactive effect of microbial inoculants on agronomical, physiological and biochemical aspects of Crocus growth, corm multiplication and apo-carotenoid biosynthesis. Significant differences in nutrient mobilization potential, induced systemic resistance, plant growth, corm, and stigma yield/quality were detected between the different biofertilizer combinations. The arbuscular mycorrhizal fungi (AMF) + phosphate-solubilizing bacteria (PSB) + potassium-solubilizing bacteria (KSB) combination was the most suitable for enhancing shoot biomass, root depth, and root biomass. Additionally, the average number of flowers per corm, stigma yield, and corm yield were greatest for the same combination. The highest multiplication rate of mother corms and a small corm index were recorded for plants inoculated with Azospirillum + PSB + KSB. Although Azospirillum caused maximum multiplication, the percentage of smaller daughter corms was greater, while the replacement of Azospirillum with AMF resulted in maximum percentage of larger corms. Plant health improved through AMF-mediated microbial interactions, which induced systemic resistance via enzymatic and nonenzymatic mechanisms and increased the uptake of P, K, Cu, Fe, Mn, and Zn. The interrelationship between plant and microbial consortium was complex, causing trait-specific qualitative and quantitative impacts on saffron attributes. Microbial inoculants increased nutrient uptake by saffron plants and triggered biochemical defence response by causing mild stress, making the plant more growth-responsive and better equipped for defence. Microbial interaction increased saffron yield and quality by favourably modulating plant metabolism, majorly overlapping with plant protection mechanisms. However, plant-microbial combinations in an organic production module are trait-specific and should be chosen carefully.

The Six-Transmembrane Enzyme GDE2 Is Required for the Release of Molecularly Distinct Small Extracellular Vesicles from Neurons.

Extracellular vesicles (EVs) are implicated in a multitude of physiological and pathophysiological processes in the nervous system; however, their biogenesis and cargoes are not well defined. Glycerophosphodiester Phosphodiesterase 2 (GDE2 or GDPD5) is a six-transmembrane protein that cleaves the Glycosylphosphatidylinositol (GPI)-anchor that tethers some proteins to the membrane and has important roles in neurodevelopment and disease-relevant pathways of neuronal survival. We show here that GDE2 regulates the number of small EVs (sEVs) released from the cell surface of neurons via its GPI-anchor cleavage activity and contributes to the loading of protein cargo through enzymatic and non-enzymatic mechanisms. Proteomic profiling reveals that GDE2 releases at least two distinct EV populations, one containing GDE2 itself and the other harboring the putative ectosomal markers CD9 and BSG. sEVs released by GDE2 are enriched in cytoskeletal and actin-remodeling proteins, suggesting a potential mechanism for GDE2-dependent EV release. Further, sEV populations released by GDE2 are enriched in proteins responsible for modulating synaptic activity and proteins that are critical for cellular redox homeostasis. These studies identify GDE2 as a novel regulator of molecularly distinct sEV populations from neurons with potential roles in the synaptic and redox pathways required for neuronal function and survival.

Spatial transcriptomic validation of a biomimetic model of fibrosis enables re-evaluation of a therapeutic antibody targeting LOXL2

Matrix stiffening by lysyl oxidase-like 2 (LOXL2)-mediated collagen cross-linking is proposed as a core feedforward mechanism that promotes fibrogenesis. Failure in clinical trials of simtuzumab (the humanized version of AB0023, a monoclonal antibody against human LOXL2) suggested that targeting LOXL2 may not have disease relevance; however, target engagement was not directly evaluated. We compare the spatial transcriptome of active human lung fibrogenesis sites with different human cell culture models to identify a disease-relevant model. Within the selected model, we then evaluate AB0023, identifying that it does not inhibit collagen cross-linking or reduce tissue stiffness, nor does it inhibit LOXL2 catalytic activity. In contrast, it does potently inhibit angiogenesis consistent with an alternative, non-enzymatic mechanism of action. Thus, AB0023 is anti-angiogenic but does not inhibit LOXL2 catalytic activity, collagen cross-linking, or tissue stiffening. These findings have implications for the interpretation of the lack of efficacy of simtuzumab in clinical trials of fibrotic diseases.

Aflatoxin B1 Control by Various Pseudomonas Isolates.

The climate-change-coupled fungal burden in crop management and the need to reduce chemical pesticide usage highlight the importance of finding sustainable ways to control Aspergillus flavus. This study examines the effectiveness of 50 Pseudomonas isolates obtained from corn rhizospheres against A. flavus in both solid and liquid co-cultures. The presence and quantity of aflatoxin B1 (AFB1) and AFB1-related compounds were determined using high-performance liquid chromatography-high resolution mass spectrometry analysis. Various enzymatic- or non-enzymatic mechanisms are proposed to interpret the decrease in AFB1 production, accompanied by the accumulation of biosynthetic intermediates (11-hydroxy-O-methylsterigmatocystin, aspertoxin, 11-hydroxyaspertoxin) or degradation products (the compounds C16H10O6, C16H14O5, C18H16O7, and C19H16O8). Our finding implies the upregulation or enhanced activity of fungal oxidoreductases and laccases in response to bacterial bioactive compound(s). Furthermore, non-enzymatic reactions resulted in the formation of additional degradation products due to acid accumulation in the fermented broth. Three isolates completely inhibited AFB1 or any AFB1-related compounds without significantly affecting fungal growth. These bacterial isolates supposedly block the entire pathway for AFB1 production in the fungus during interaction. Apart from identifying effective Pseudomonas isolates as potential biocontrol agents, this work lays the foundation for exploring new bacterial bioactive compounds.

Antioxidative and Antiglycative Stress Activities of Selenoglutathione Diselenide.

The damage caused by oxidative and glycative stress to cells accumulates on a daily basis and accelerates aging. Glutathione (GSH), a major antioxidant molecule in living organisms, plays a crucial role in detoxifying the stress-causing substances inherent in cells, such as H2O2 and methylglyoxal (MG), an important intermediate of advanced glycation end-products (AGEs). In this study, we focused on the enhanced antioxidant capacity of the selenium analog of GSH, i.e., selenoglutathione (GSeH), compared to GSH, and examined its effects on the detoxification of stress-causing substances and improvement in cell viability. In cell-free systems, GSeH (1 mM) generated in situ from GSeSeG in the presence of NADPH and glutathione reductase (GR) rapidly reduced more than 80% of 0.1 mM H2O2, indicating the significant glutathione peroxidase (GPx)-like antioxidant activity of GSeSeG. Similarly, around 50% of 0.5 mM MG was degraded by 0.5 mM GSeH within 30 min through a non-enzymatic mechanism. It was also found that GSeSeG (0.05-0.5 mM) showed glutathione S-transferase (GST)-like activity against 1-chloro-2,4-dinitrobenzene (CDNB), a model substance of oxidative stress-causing toxic materials in cells. Meanwhile, HeLa cells that had been pre-treated with GSeSeG exhibited increased viability against 1.2 mM H2O2 (at [GSeSeG] = 0.5-50 μM) and 4 mM MG (at [GSeSeG] = 3 μM), and the latter effect was maintained for two days. Thus, GSeSeG is a potential antioxidant and antiglycative stress agent for cells.

Heavy Metal Stress and Cellular Antioxidant Systems of Plants: A Review

Heavy metal (HM) stress is one of the most important abiotic stresses affecting flora and fauna worldwide due to a rapid global increase in the urbanization and industrialization and the consequent rise of HM concentration in the atmosphere. Typically, plants have distinct mechanisms to cope up with HM stress. These mechanisms rendering tolerance to the plants by detoxifying the HMs. Additionally, a number of physiological and molecular changes occur in plant cells due to their exposure to HMs. This culminates in the generation of reactive oxygen species (ROS) that cause oxidative stress in plants, which significantly affects plant metabolism and disrupts normal vital cellular functions. However, in order to cope with such stresses, plants possess a strong antioxidant defence system to counteract increases in the ROS-induced stress. The enzymatic components of this system include superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX), monodehydroascorbate reductase (MDHAR), dehydroascorbate reductase (DHAR) and glutathione reductase (GR). The non-enzymatic components are ascorbate, glutathione and phenolic compounds along with lipid-soluble molecules such as carotenoids and tocopherols. This review makes an effort to collect and collate the currently available information on metal stress and cellular antioxidant systems of plants, emphasizing upon the role of enzymatic and non-enzymatic mechanisms for detoxification of HM-induced oxidative stress.

Preparation of 2-Aminoimidazole-Activated Substrates for the Study of Nonenzymatic Genome Replication.

Nonenzymatic genome replication is thought to be an important process for primitive lifeforms, but this has yet to be demonstrated experimentally. Recent studies on the nonenzymatic primer extension mechanism mediated by nucleoside 5'-monophosphates (NMPs) activated with 2-aminoimidazole have revealed that imidazolium-bridged dinucleotide intermediates (N*N) account for the majority of the chemical copying process. As a result, an efficacious synthetic pathway for producing substrates activated with an imidazoyl moiety is desirable. This article provides a detailed protocol for the standard dehydrative redox reaction between NMPs and 2-aminoimidazole to produce nucleotide phosphoroimidazolides. In addition, we describe a similar synthetic pathway to produce N*N in high yields for homodimers. Finally, a simple reversed-phase cation exchange step is described to increase NMP solubility, which significantly increases yields for certain substrates. This approach allows for an efficient and cost-effective methodology to prepare high-quality substrates utilized in origins-of-life studies. © 2024 The Author(s). Current Protocols published by Wiley Periodicals LLC. Basic Protocol 1: Synthesis of 2-aminoimidazolephosphoroimidazolide-activated cytidine Basic Protocol 2: Synthesis of 2-aminoimidazolium-bridged dicytidyl intermediate Basic Protocol 3: Cation exchange of guanosine 5'-monophosphate disodium salt Alternate Protocol: Synthesis of cytidine 5'-phosphoroimidazolide or 2-aminoimidazolium-bridged dicytidyl from cytidine 5'-monophosphate disodium salt.

Functionality of Reactive Oxygen Species (ROS) in Plants: Toxicity and Control in Poaceae Crops Exposed to Abiotic Stress.

Agriculture and changing environmental conditions are closely related, as weather changes could adversely affect living organisms or regions of crop cultivation. Changing environmental conditions trigger different abiotic stresses, which ultimately cause the accumulation of reactive oxygen species (ROS) in plants. Common ROS production sites are the chloroplast, endoplasmic reticulum, plasma membrane, mitochondria, peroxisomes, etc. The imbalance in ROS production and ROS detoxification in plant cells leads to oxidative damage to biomolecules such as lipids, nucleic acids, and proteins. At low concentrations, ROS initiates signaling events related to development and adaptations to abiotic stress in plants by inducing signal transduction pathways. In plants, a stress signal is perceived by various receptors that induce a signal transduction pathway that activates numerous signaling networks, which disrupt gene expression, impair the diversity of kinase/phosphatase signaling cascades that manage the stress response in the plant, and result in changes in physiological responses under various stresses. ROS production also regulates ABA-dependent and ABA-independent pathways to mitigate drought stress. This review focuses on the common subcellular location of manufacturing, complex signaling mechanisms, and networks of ROS, with an emphasis on cellular effects and enzymatic and non-enzymatic antioxidant scavenging mechanisms of ROS in Poaceae crops against drought stress and how the manipulation of ROS regulates stress tolerance in plants. Understanding ROS systems in plants could help to create innovative strategies to evolve paths of cell protection against the negative effects of excessive ROS in attempts to improve crop productivity in adverse environments.

Using seasonal physiological and biochemical responses to select forest components adapted to soybean and corn intercropping

Given the increasing utilization of forest components in integration systems worldwide, coupled with the growing demand for food in regions facing water restrictions, this study aims to evaluate how physiological and biochemical parameters contribute to the diversification of adaptive mechanisms among native species and eucalyptus genotypes intercropped with soybean or corn. The native tree species Anadenanthera macrocarpa and Dipteryx alata, and the eucalyptus genotypes Urograndis I-144 and Urocam VM01, were grown in soybean and corn intercropping areas and evaluated in fall, winter, spring, and summer. The study evaluated morning water potential, chloroplast pigment concentration, gas exchange, cell damage, and antioxidant enzyme activity. Intercropped with soybean, development the of A. macrocarpa improved through instantaneous water use efficiency, energy use by the electron transport chain, chloroplast pigments, and catalase enzyme activity. On the other hand, A. macrocarpa when, intercropped with corn, despite increasing energy absorption by the reaction center, there is a need for non-photochemical dissipation and in the activity of the enzymes superoxide dismutase and ascorbate peroxidase in response to water and oxidative deficits. In D. alata, the physiological and biochemical responses were not influenced by intercropping but by seasons, with increased chloroplast pigments in fall and electron transport in summer. However, in corn intercropping, the dissipation of excess energy allowed leaf acclimatization. The I-144 and VM01 genotypes also showed no significant differences between intercrops. The results describe photosynthetic and biochemical challenges in the native species A. macrocarpa intercropped with corn, such as a greater need for enzymatic and non-enzymatic defense mechanisms in response to more negative water potential. In D. alata, the challenges are present in both intercrops due to improved mechanisms to protect the photosynthetic apparatus. The survival of the I-144 genotype may be inefficient in both intercrops under prolonged drought conditions, as it modifies the photosystem; in contrast, genotype VM01 was the most adapted to the system for using captured energy, reducing water loss and being resilient.

Regulation of nitro-oxidative homeostasis: an effective approach to enhance salinity tolerance in plants.

Soil salinity is a major constraint for sustainable agricultural productivity, which together with the incessant climate change may be transformed into a severe threat to the global food security. It is, therefore, a serious concern that needs to be addressed expeditiously. The overproduction and accumulation of reactive oxygen species (ROS) and reactive nitrogen species (RNS) are the key events occurring during salt stress, consequently employing nitro-oxidative stress and programmed cell death in plants. However, very sporadic studies have been performed concerning different aspects of nitro-oxidative stress in plants under salinity stress. The ability of plants to tolerate salinity is associated with their ability to maintain the cellular redox equilibrium mediated by both non-enzymatic and enzymatic antioxidant defense mechanisms. The present review emphasizes the mechanisms of ROS and RNS generation in plants, providing a detailed evaluation of how redox homeostasis is conserved through their effective removal. The uniqueness of this article stems from its incorporation of expression analyses of candidate genes for different antioxidant enzymes involved in ROS and RNS detoxification across various developmental stages and tissues of rice, utilizing publicly available microarray data. It underscores the utilization of modern biotechnological methods to improve salinity tolerance in crops, employing different antioxidants as markers. The review also explores how various transcription factors contribute to plants' ability to tolerate salinity by either activating or repressing the expression of stress-responsive genes. In summary, the review offers a thorough insight into the nitro-oxidative homeostasis strategy for extenuating salinity stress in plants.

High Burden of Carbapenem-Resistant Organisms Screened by Xpert Carba-R Assay Versus Culture from Intensive Care Patients in Indonesia

The World Health Organization (WHO) considers carbapenem-resistant organisms (CROs) to be critical-level pathogens. Regular screening for high-risk CRO colonization is essential, especially in the ICU. Direct detection of carbapenem-resistant genes is possible using the FDA-approved Xpert Carba-R assay. This study evaluated its reliability compared with the culture technique at a tertiary hospital in Indonesia. A high number of CRO colonization was found using the culture technique and the Xpert Carba-R assay with about 31 and 26 positive results out of 100 total samples, respectively. Both methods detected blaNDM in 11 samples, and the Xpert Carba-R assay detected one sample co-presenting with blaVIM that was not detected by PCR. The Xpert Carba-R assay did not detect the gene in 73 samples following negative results with the culture technique. Fifteen samples were detected gene by the Xpert Carba-R assay though there was no gene by the culture method, showing that the Xpert Carba-R assay demonstrated a high degree of sensitivity in identifying carbapenem-resistance genes. Carbapenem-resistance genes common in Indonesia other than those examined by Xpert Carba-R assay in this study (i.e., blaOXA-23 and blaOXA-24) or non-enzymatic mechanisms may also produce resistance in many colonies without the examined genes. Finally, the Xpert Carba-R assay produced faster findings than the culture technique.

Evolution of mutations in the ftsI gene leading to amino acid substitutions in PBP3 in Haemophilus influenzae strains under the selective pressure of ampicillin and cefuroxime

BackgroundAminopenicillins are recommended agents for non-invasive Haemophilus influenzae infections. One of the mechanisms of resistance to β-lactams is the alteration of the transpeptidase region of penicillin binding protein 3 (PBP3) which is caused by mutations in the ftsI gene. It was shown that exposure to beta-lactams has a stimulating effect on increase of prevalence of H. influenzae strains with the non-enzymatic mechanism of resistance. ObjectivesThe aim of our study was to compare the mutational potential of ampicillin and cefuroxime in H. influenzae strains, determination of minimum inhibitory concentration and the evolution of mutations over time, focusing on amino acid substitutions in PBP3. Methods30 days of serial passaging of strains in liquid broth containing increasing concentrations of ampicillin or cefuroxime was followed by whole-genome sequencing. ResultsOn average, cefuroxime increased the minimum inhibitory concentration more than ampicillin. The minimum inhibitory concentration was increased by a maximum of 32 fold. Substitutions in the PBP3 started to appear after 15 days of passaging. In PBP3, cefuroxime caused different substitutions than ampicillin. ConclusionsOur experiment observed differences in mutation selection by ampicillin and cefuroxime. Selection pressure of antibiotics in vitro generated substitutions that do not occur in clinical strains in the Czech Republic.